Effect Of Different Warm Ischemia Time On Orthotopic Liver Transplantation Of Cardiac Death Moderate Fatty Liver Rats

【Objective】1. To establish a stable, reliable, reproducible and inexpensive rat model of moderatenon-alcoholic fatty liver.2. To establish a stable, reliable and reproducible rat model of DCD fatty liver fortransplantation.3. To explore effect of different warm ischemia time(WIT0,WIT10,WIT20) on DCDfatty liver rats after liver transplantation from survival rate, function andhistopathological examination.【Methods】1. The first experiment: Establishment and observation of moderate non-alcoholicfatty liver rat model:1.1Animal groups:20rats were randomly divided into high-fat diet group and thenormal diet group, each group was10rats, high-fat diet group was fed high fat diet,normal diet group was fed a normal diet.1.2Moderate non-alcoholic fatty liver rat model: feeding with high-fat diet (formulais88%basal diet,10%lard and2%cholesterol) for6weeks.1.3Observation: After6weeks, to compare the following data with two groups: bodyweight, liver wet weight, liver index, liver function and degree of fatty liver accordingto histopathological examination.2. The second experiment: Establishment and observation of DCD fatty livertransplantation rat model:2.1DCD model: Thoracotomy-induced cardiac death, cardiac arrest as a start ofwarm ischemia time; to monitor arterial blood pressure when we establish DCD rat model though femoral artery cannulation.2.2Orthotopic liver transplantation model: Refer to Kamada’s two-cuff method.2.3Observation: Arterial blood pressure and time from thoracotomy to cardiac death;10cases of DCD moderate fatty liver rats were completed transplantation successfully,warm ischemia time is5min, calculate success rate of modeling.3. The third experiment: To explore the warm ischemia time of DCD moderate fattyliver rats for safe transplantation:3.1Animal groups: according to warm ischemia time, rats were divided into3groups(WIT0, WIT10, WIT20),51cases finished DCD fatty liver transplantation foreach group.3.2Observation:1d,3d,5d,7d, and14d after transplantation each group killed6ratsrandomly, then compare liver function, liver pathology and30days survival rate.4. Statistical analysis: All data were analyzed by SPSS13.0statistical software. Allmeasurement data were presented as mean±SD (x s). Using two-sample t test,ANOVA+LSD method, factorial analysis of variance, repeated measures analysisof ANOVA to analyze all measurement data. Kaplan-Meier analysis and log-ranktest were used to analyze survival rates and difference among various groups. α=0.05was considered significant.【Results】1. Establishment and observation of moderate non-alcoholic fatty liver rat model:1.1Compared with normal diet group, AST and ALT of the high-fat diet group wasslightly higher, TG was significantly higher. Both had statistical significance(P <0.05).1.2There were no difference between two groups for body weight, liver wet weightand liver index (P<0.05);1.3Feeding with high-fat diet for6weeks can copy non-alcoholic moderate fatty liverrat model successfully, success rate was100%;2. Establishment and observation of DCD fatty liver transplantation rat model:2.1Arterial blood pressure: Within1min after thoracotomy, donor rats showed atransient increase in systolic blood pressure due to the suffocation. During this period,systolic pressure increased from (94.70±6.45) mmHg to (108.70±7.42) mmHg (P <0.05), then decreased rapidly to0mmHg when cardiac arrest.2.2Cardiac arrest time:8.6±2.1min after thoracotomy can induce cardiac arrest.2.310cases of DCD moderate fatty liver rats were completed transplantationsuccessfully, one rat died of SHVC anastomotic bleeding3h after surgery, success ratewas90%(9/10).3. To explore the warm ischemia time of DCD moderate fatty liver rats for safetransplantation:3.1There were no difference for30d survival rate between WIT0group and WIT10group (P>0.05). All15rats of WIT20group died of PNF within2days after surgery,the survival rate was significantly lower than other groups (P <0.05).3.2Blood index: After operation, serum AST and ALT of WIT10group on1d,3d,5d,7d,14d were higher than WIT0group(P <0.05). TBIL, ALB and PT were nosignificant difference (P>0.05) in each time point. Compared with WIT0group andWIT10group,AST, ALT, TBIL and PT in WIT20group were significantly higher on1d after surgery (P<0.05), But ALB was significantly lower (P <0.05).3.3HE staining results:With the extension of warm ischemia time， liverHistopathological damage was more serious. Within10min, liver damage wasreversible, But over20min, irreversible damage such as large areas of necrosishappened.【Conclusion】1. High-fat diet can induce rat model of moderate non-alcoholic fatty liver.It isinexpensive, simple and reproducible.2. The rats of Thoracotomy-induced cardiac death and Kamada’s two-cuff method forDCD fatty liver transplantation is ideal small animal models for simulating clinicalDCD fatty liver transplantation.3. Fatty liver graft is sensitive to warm and ischemia. With the extension of warmischemia time, liver function is worse and liver Histopathological damage is moreserious. When the warm ischemia time is over10minutes, survival rate ofrats decrease significantly. Within10minutes is the warm ischemia time of DCDmoderate fatty liver rats liver for safe transplantation.